喜树叶、枝水溶性糖提取的研究

用正交实验结果作方差分析表明：溶剂用量对试验结果有较显著的影响（a=0.10)，提取温度对试验结果也有较大的影响，而提取时间对试验结果的影响不显著。喜树叶中水溶性糖的提取最优方案是A3C2B3，即溶剂用量为60ml，提取温度80℃，提取时间60min；喜树叶水溶性糖的提取最优方案是A2C2B1，即溶剂用量为40ml，提取温度80℃，提取时间20min。通过对喜树叶与枝的成对比较分析，叶与枝的水溶性糖含量无显著差异。     

Synthesis of 3′,4′-difluoro-3′-deoxyribonucleosides and its evaluation of the biological activities: Discovery of a novel type of anti-HCV agent 3′,4′-difluorocordycepin

Upon reacting 3′,4′-unsaturated cytosine (8 and 9) and adenine nucleosides (13 and 14) with XeF₂/BF₃·OEt₂, the respective novel 3′,4′-difluoro-3′-deoxyribofuranosyl nucleosides (10–12 and 15–18) could be obtained. Formation of anti-adducts (11, 16 and 18) revealed that the fluorination involved oxonium ions as incipient intermediates. TBDMS-protected 3′,4′-unsaturated adenosine provided the β-face adducts as sole stereoisomers whereas α-face-selectivity was observed with the TBDPS-protected adenosine 14. The evaluation of the novel 3′-deoxy-3′,4′-difluororibofuranosylcytosine-(19–21) and adenine nucleosides (22–25) against antitumor and antiviral activities revealed that 3′,4′-difluorocordycepin (24) was found to possess anti-HCV activity. The SI of 24 was comparable to that of the anti-HCV drug ribavirin. However, sofosbuvir, FDA-approved novel anti-HCV drug, showed better SI value. Our finding revealed that the introduction of the fluoro-substituent into the 4′-position of cordycepin derivatives decreased the cytotoxicity to the host cell with retention of the antiviral activity.     

Dinitrogen fixation and infection of grass leaves byPseudomonas rubrisubalbicans andHerbaspirillum seropedicae

Bacteria causing mottled stripe disease in sugar cane, known asPseudomonas rubrisubalbicans, were shown to be able to fix molecular N₂ and to grow on it. The root associated diazotroph known asHerbaspirillum seropedicae, after artificial inoculation caused mottled stripe disease symptoms on sorghum and Napier grass but not on sugar cane. Both bacteria could be reisolated from leaves even 60 days after. Sugar cane leaves contained large numbers of these bacteria even in the uninoculated controls. Additional physiological characteristics of six strains ofP. rubrisubalbicans were compared with those of twoH. seropedicae strains and were shown to be very similar.     

Significant impact of divalent metal ions on the fidelity,sugar selectivity,and drug incorporation efficiency of human PrimPol

Human PrimPol is a recently discovered bifunctional enzyme that displays DNA template-directed primase and polymerase activities. PrimPol has been implicated in nuclear and mitochondrial DNA replication fork progression and restart as well as DNA lesion bypass. Published evidence suggests that PrimPol is a Mn²⁺-dependent enzyme as it shows significantly improved primase and polymerase activities when binding Mn²⁺, rather than Mg²⁺, as a divalent metal ion cofactor. Consistently, our fluorescence anisotropy assays determined that PrimPol binds to a primer/template DNA substrate with affinities of 29 and 979 nM in the presence of Mn²⁺ and Mg²⁺, respectively. Our pre-steady-state kinetic analysis revealed that PrimPol incorporates correct dNTPs with 100-fold higher efficiency with Mn²⁺ than with Mg²⁺. Notably, the substitution fidelity of PrimPol in the presence of Mn²⁺ was determined to be in the range of 3.4 × 10⁻² to 3.8 × 10⁻¹, indicating that PrimPol is an error-prone polymerase. Furthermore, we kinetically determined the sugar selectivity of PrimPol to be 57–1800 with Mn²⁺ and 150–4500 with Mg²⁺, and found that PrimPol was able to incorporate the triphosphates of two anticancer drugs (cytarabine and gemcitabine), but not two antiviral drugs (emtricitabine and lamivudine).     

Sugar‐feeding status alters biting midge photoattraction

The biting midge Culicoides sonorensis Wirth and Jones (Diptera: Ceratopogonidae) transmits pathogens to both livestock and wildlife. Biting midge surveillance relies heavily on light traps for collection; however, little is known about the light spectra preferences of C. sonorensis midges. A light assay arena was constructed and light‐emitting diodes (LEDs) of various light spectra were used as light sources to evaluate midge photoattraction. A comparison of responses to light spectra indicated the highest proportions of C. sonorensis were attracted to ultraviolet (UV) light and that midges differentiated 10‐nm differences in wavelength. Stronger intensities of UV light resulted in greater attraction. Midges exhibited both sugar‐seeking and escape behaviours under different conditions of sugar supplementation before and during the experiment. These behaviours occurred with lights of 355 nm and 365 nm in wavelength. Based on the results of this study, the attraction of C. sonorensis to light traps can be improved through the use of bright LEDs at 355 nm or 365 nm.     

Relationship between the concentration of sugars in the roots and VA mycorrhizal infection

Summary The VA-infected wheat varieties showed an increase of total (Lozano var.) and reducing (Lozano and Pane vars.) sugars in their root extracts. However, no clear relationship between sugar concentration in the root and VA mycorrhizal infection level could be established.In addition, the VA mycorrhizal hosts sorghum, alfalfa, sunflower and maize, and non-host radish and cabbage plants were tested for sugar content in their root extracts after fifteen days of growth. Sugars present in the root extracts of these plants did not seem to be a decisive factor in plant susceptibility to VA infection.     

Arg-Gly-Asp (RGD) sequence conjugated in a synthetic copolymer bearing a sugar moiety for improved culture of parenchymal cells (hepatocytes)

A copolymer, including a Gly-Arg-Gly-Asp-Ser (GRGDS) sequence and sugar moieties, was synthesized for the culturing of parenchymal cells (hepatocytes). Hepatocyte cells attached to poly[N-p-vinylbenzyl-d-maltonamide-co-6-(p-vinylbenzamido)-hexanoic acid-GRGDS] [poly(VMA-co-VBRGD)]-coated dishes grew approximately 60% better than on other polymer-coated surface for 12 h. Also, about 80% greater albumin secretion (0.38 pg ml^–1) and about 70% greater urea synthesis (0.495 pg ml^–1) from hepatocytes were produced in this matrix as compared with unstimulated cells. The behaviour of hepatocytes on poly(VMA-co-VBGRGDS)-coated dishes was not distinct from those attached to a collagen. The conjugation of the adhesion molecules of the RGD peptide in the poly(VMA-co-VBGRGDS) copolymer therefore specifically interacts with integrin families on the hepatocyte cell membrane.     

Influence of body size on fecundity and sperm management in the parasitoid wasp Anisopteromalus calandrae

A large body size is considered to be advantageous to the reproductive success of females as a result of several factors, such as the allocation of more resources to reproduction and the efficient management of sperm transferred by males. In the present study, the effects of female body size, female mating status and additional food availability on fecundity and the offspring sex ratio are investigated in the parasitoid wasp Anisopteromalus calandrae Howard (Hymenoptera: Pteromalidae). Because of haplodiploid sex determination, females must fertilize eggs to produce female offspring but not to produce male offspring. As predicted, female fecundity and the number of female offspring are positively correlated with body size. However, although the volume of the spermatheca increases with female body size, the amount of sperm stored in the spermatheca is relatively constant, irrespective of body size. Consequently, larger females produce a greater proportion of male offspring, especially at the end of the oviposition sequence, suggesting that larger females that possess more resources for reproduction and produce a larger number of offspring are more likely to suffer sperm depletion. The results of the present study also show that mated females have an increased fecundity compared with virgin females, although the opportunity to feed on honey along with host feeding has no impact upon fecundity or the sex ratio.     

模拟根系分泌物输入对高寒退化草地土壤微生物残体的影响

微生物残体是稳定土壤碳库的重要来源，对退化生境碳的固持和积累具有重要意义。植物根系分泌物作为植物-土壤-微生物"交流"的媒介，是调控土壤微生物残体迁移转化的关键。因此，以极度退化草地土壤为对象，以氨基糖为标志物，模拟研究了不同氮浓度（低氮-LN：0.1 gN/kg；高氮-HN：0.2 gN/kg）和多样性（3种化合物、9种化合物）根系分泌物输入对土壤微生物残体的影响。结果表明：（1）根系分泌物输入可显著增加高寒退化草地土壤微生物残体含量，且主要由真菌残体贡献。其中高氮和低多样性处理增加最明显，微生物残体和真菌残体分别增加了101.14%，125.16%，而低氮和高多样性处理微生物残体和真菌残体仅增加了35.79%，33.51%。（2）根系分泌物的输入可增加土壤β-葡萄糖苷酶、土壤磷酸酶和过氧化物酶活性，促进微生物的生长，而降低β-N-乙酰氨基葡萄糖苷酶活性，减少微生物残体的分解。（3）回归分析结果显示，土壤微生物残体与土壤环境的C/N呈显著负相关，与微生物生物量C/N呈显著正相关。上述结果表明，在未来退化草地恢复中，可充分利用模拟根系分泌物输入的土壤固碳策略，即通过提高土壤氮的有效性，促进微生物的生长，加快代谢周转，进一步提高微生物残体含量。